10CFU™ Mycoplasma Sensitivity Standards

Mycoplasma tests according to the European Pharmacopoeia (EP) Chapter 2.6.7 require a sensitivity of 10 colony forming units (CFU) per ml sample volume for NAT-based methods such as PCR in order to replace traditional culture methods. This sensitivity must be accomplished as part of the robustness testing for any particular sample matrix.

Using vital mycoplasma is not acceptable for the majority of cell culture laboratories due to safety regulations. Our 10CFU™ sensitivity standards contain non-vital material and allow safe and reliable validation.

The mycoplasma were grown in culture medium as described in EP 2.6.7, subsequently titrated and plated on Hayflick and Frey medium for CFU determination. The mycoplasma were harvested in the early logarithmic growth phase to ensure a high ratio of vital to non-vital mycoplasma and thus a low ratio of GU* to CFU. All strains were obtained from the National Collection of Type Cultures (NCTC), UK, and cultivated in low passages.

Scope of Application

10CFU™ Sensitivity Standards are intended for validating the robustness and sensitivity of NAT-based mycoplasma tests.

Kit Components

Unit package: 3 vials containing 10 cfu of the particular mycoplasma as well as 3 vials negative control

Set package: 2 vials of each EP2.6.7-listed mycoplasma containing 10 cfu (18 vials in total) as well as 2 vials negative control

The components are lyophilized.

Species and Order Information

  • Cat. No. 102-1003   Mycoplasma arginini
  • Cat. No. 102-2003   Mycoplasma orale 
  • Cat. No. 102-3003   Mycoplasma gallisepticum
  • Cat. No. 102-4003   Mycoplasma pneumoniae
  • Cat. No. 102-5003   Mycoplasma synoviae
  • Cat. No. 102-6003   Mycoplasma fermentans
  • Cat. No. 102-7003   Mycoplasma hyorhinis
  • Cat. No. 102-8003   Acholeplasma laidlawii
  • Cat. No. 102-9003   Spiroplasma citri
  • Cat. No. 102-0002   Mycoplasma Set


Application of a PCR-based mycoplasma test, e.g. Microsart® ATMP Mycoplasma kit

Required Consumables

Sterile 1.5 ml reaction tubes, filter tips, appropriate DNA extraction kit

Required lab devices

Microcentrifuge, micropipettes

Shelf Life and Storage

The date of expiry is given on each package. The unopened kit should be stored at +2 to +8 °C.

Figure - 10CFU were re-suspended in 1 ml cell culture medium, 200 µl were used for DNA extraction with the Venor®GeM Sample Preparation Kit. The mycoplasma PCR-test was performed with the Microsart® ATMP Mycoplasma kit and a Corbett RotorGene®6000 cycler.